ABSTRACT
Mycobacterium tuberculosis remains a leading cause of death worldwide, especially among individuals infected with HIV. Whereas phylogenetic analysis has revealed M. tuberculosis spread throughout history and in local outbreaks, much less is understood about its dissemination within the body. Here we report genomic analysis of 2,693 samples collected post mortem from lung and extrapulmonary biopsies of 44 subjects in KwaZulu-Natal, South Africa, who received minimal antitubercular treatment and most of whom were HIV seropositive. We found that purifying selection occurred within individual patients, without the need for patient-to-patient transmission. Despite negative selection, mycobacteria diversified within individuals to form sublineages that co-existed for years. These sublineages, as well as distinct strains from mixed infections, were differentially distributed throughout the lung, suggesting temporary barriers to pathogen migration. As a consequence, samples taken from the upper airway often captured only a fraction of the population diversity, challenging current methods of outbreak tracing and resistance diagnostics. Phylogenetic analysis indicated that dissemination from the lungs to extrapulmonary sites was as frequent as between lung sites, supporting the idea of similar migration routes within and between organs, at least in subjects with HIV. Genomic diversity therefore provides a record of pathogen diversification and repeated dissemination across the body.
Subject(s)
DNA, Bacterial/genetics , HIV Infections/complications , Liver/microbiology , Lung/microbiology , Lymph Nodes/microbiology , Mycobacterium tuberculosis/genetics , Spleen/microbiology , Tuberculosis/microbiology , Adult , Aged , Autopsy , Bacteriological Techniques , Coinfection/microbiology , Female , Genetic Variation , Humans , Male , Middle Aged , Phylogeny , Polymorphism, Single Nucleotide , South Africa , Tuberculosis/complications , Tuberculosis, Hepatic/complications , Tuberculosis, Hepatic/microbiology , Tuberculosis, Lymph Node/complications , Tuberculosis, Lymph Node/microbiology , Tuberculosis, Pulmonary/complications , Tuberculosis, Pulmonary/microbiology , Tuberculosis, Splenic/complications , Tuberculosis, Splenic/microbiologyABSTRACT
The clinical management of tuberculosis is a major challenge in southern Africa. The prevalence of within-host genetically heterogeneous Mycobacterium tuberculosis infection and its effect on treatment response are not well understood. We enrolled 500 patients with tuberculosis in KwaZulu-Natal and followed them through 2 months of treatment. Using mycobacterial interspersed repetitive units-variable number of tandem repeats genotyping to identify mycobacterial heterogeneity, we report the prevalence and evaluate the association of heterogeneity with treatment response. Upon initiation of treatment, 21.1% of participants harbored a heterogeneous M. tuberculosis infection; such heterogeneity was independently associated with a nearly 2-fold higher odds of persistent culture positivity after 2 months of treatment (adjusted odds ratio, 1.90; 95% confidence interval, 1.03-3.50).
